Single and few cell analysis for correlative light microscopy, metabolomics, and targeted proteomics
Luca Rima1, Christian Berchtold2, Stefan Arnold1
1Biozentrum, University of Basel, Spitalstrasse 41, Basel, Switzerland.
Lab on a Chip
|August 12, 2024
Summary
This study introduces a new method to analyze metabolites and proteins in single eukaryotic cells. This technique enables correlative measurements of cellular structures, metabolites, and proteins at the single-cell level.
Area of Science:
- Cell Biology
- Biochemistry
- Analytical Chemistry
Background:
- Cellular phenotypes are shaped by complex interactions of biomolecules.
- Stochastic cellular processes and individual cell development hinder population synchronization.
- Single-cell analysis is crucial to overcome signal dilution and averaging effects.
Purpose of the Study:
- To develop a method for analyzing metabolites and proteins from a single or few adherent eukaryotic cells.
- To enable correlative measurements of cellular components at the single-cell level.
Main Methods:
- Cells are lysed via electroporation and aspirated using a microcapillary.
- Lysates are analyzed using liquid-chromatography mass spectrometry (LC-MS) for metabolites.
- Reverse-phase protein arrays (RPPA) are used for targeted protein detection.
- Correlative measurements integrate light microscopy, LC-MS, and RPPA.
Main Results:
- A method was established to analyze metabolites and proteins from single or few cells.
- Correlative measurements of cellular structure, metabolites, and proteins were achieved on the few-cell level.
- The technique allows for simultaneous assessment of cellular structures, metabolites, and targeted proteins.
Conclusions:
- The developed method facilitates single-cell analysis of metabolites and proteins.
- This approach enables correlative studies of cellular components, advancing understanding of cell heterogeneity.
- Potential applications and limitations for future improvements were discussed.
